Proteomics reveals time-dependent protein corona changes in the intracellular pathway

Area of Science:

• Nanotechnology-biology interactions
• Cellular nanotechnology
• Nanoparticle-protein interactions

Background:

• The intracellular protein corona, proteins coating nanoparticles within cells, remains understudied.
• Understanding its formation and dynamics is vital for nanocarrier efficacy in drug delivery.
• Most nanocarriers are designed for cellular uptake, necessitating intracellular interaction studies.

Purpose of the Study:

• To establish a workflow for isolating and analyzing the intracellular protein corona.
• To investigate the formation and dynamics of intracellular protein coronas for two distinct nanoparticles.
• To correlate intracellular protein corona composition with nanocarrier uptake kinetics and trafficking pathways.

Main Methods:

• Workflow development for intracellular protein corona isolation.
• Label-free quantitative LC-MS proteomics for proteomic analysis.
• Flow cytometry, TEM, and confocal microscopy for validation and trafficking studies.

Main Results:

• Demonstrated distinct intracellular protein corona evolution for nanoparticles with different uptake rates (HES-NPs vs. HSA-NCs).
• Identified key proteins involved in macropinocytosis and caveolin-mediated uptake.
• Confirmed predominant trafficking from early endosomes to late endosomes/lysosomes.

Conclusions:

• Analysis of the intracellular protein corona provides critical data for understanding nanocarrier trafficking.
• Differences in nanoparticle uptake kinetics significantly influence intracellular protein corona composition.
• This research advances the understanding of intracellular nanocarrier behavior to improve drug delivery systems.